DOI: 10.1073/pnas.96.1.91. Gene delivery: A single nuclear localization signal peptide is sufficient to carry DNA to the cell nucleus

Gene delivery: A single nuclear localization signal peptide is sufficient to carry DNA to the cell nucleus

10.1073/pnas.96.1.91

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Translocation of exogenous DNA through the nuclear membrane is a major concern of gene delivery technologies. To take advantage of the cellular import machinery, we have synthesized a capped 3.3-kbp CMVLuciferase-NLS gene containing a single nuclear localization signal peptide (PKKKRKVEDPYC). Transfection of cells with the tagged gene remained effective down to nanogram amounts of DNA. Transfection enhancement (10- to 1,000-fold) as a result of the signal peptide was observed irrespective of the cationic vector or the cell type used. A lysine to threonine mutation of the third NLS amino acid completely abolished these remarkable features, suggesting importin-mediated translocation. Our hypothesis is that the 3-nm-wide DNA present in the cytoplasm is initially docked to and translocated through a nuclear pore by the nuclear import machinery. As DNA enters the nucleus, it is quickly condensed into a chromatin-like structure, which provides a mechanism for threading the remaining worm-like molecule through the pore. A single NLS signal is thus sufficient, whereas many signals on a gene would actually inhibit entry, the same DNA molecule being threaded through adjacent pores.

Bibliography

Zanta, M. A., Belguise-Valladier, P., & Behr, J.-P. (1999). Gene delivery: A single nuclear localization signal peptide is sufficient to carry DNA to the cell nucleus. Proceedings of the National Academy of Sciences, 96(1), 91â96.

Authors 3

Maria Antonietta Zanta (first)
Pascale Belguise-Valladier (additional)
Jean-Paul Behr (additional)

References 41 Referenced 594

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Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:39 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 3:39 p.m.)
Indexed	2 weeks, 5 days ago (Aug. 7, 2025, 4:51 a.m.)
Issued	26 years, 7 months ago (Jan. 5, 1999)
Published	26 years, 7 months ago (Jan. 5, 1999)
Published Online	26 years, 7 months ago (Jan. 5, 1999)
Published Print	26 years, 7 months ago (Jan. 5, 1999)

Funders 0

None

BibTeX

@article{Zanta_1999, title={Gene delivery: A single nuclear localization signal peptide is sufficient to carry DNA to the cell nucleus}, volume={96}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.96.1.91}, DOI={10.1073/pnas.96.1.91}, number={1}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Zanta, Maria Antonietta and Belguise-Valladier, Pascale and Behr, Jean-Paul}, year={1999}, month=jan, pages={91–96} }

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